Aim:: Atherosclerosis is a kind of chronic inflammatory disease. A crucial pathology change of atherosclerosis is the migration of activated VSMCs to the intima where they interact with leukocytes by expressing adhesion molecules, including intercellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). Moreover, monocyte chemoattractant protein-1 (MCP-1) expressed by VSMCs plays an important role in recruiting monocytes and macrophages. Leech (Whitmania pigra Whitman) is a traditional Chinese medicine to treat cardiovascular diseases including atherosclerosis, however previous research has rarely reported the molecular mechanism for its curative effect. Thus, our study focuses on the effects of leech extracts on the expression of inflammatory factors, adhesion molecules and MCP-1 in rat VSMCs.

Results:: LEE suppressed LPS-induced VSMCs migration and decreased the chemotaxis and adhesive capacity of THP-1 and RAW264.7 to LPS-stimulated VSMCs. LEE also attenuated the upregulation of a variety of pro-atherosclerotic factors by inhibiting the phosphorylation of p38 MAPK. LEE was also observed to prevent NF-&kappa;B p65 nuclear localization using immune-fluorescent staining.

Figure 6: Effect of LEE on nuclear accumulation of p65 induced by LPS in VSMCsConfluent VSMCs monolayers were grown on coverslips followed by being incubated with different conditions in the 24-hole plates for 48 h. Then cells were fixed, permeabilized, and stained with anti-p65 antibody and a secondary antibody conjugated to fluorescent red. The DNA was stained using DAPI. The negative control (p65 antibody not added) and a blank control are both included.

Mentions:
The activation of NF-κB contributes to inflammation status by promoting the expression of inflammation genes. To verify NF-κB p65 nuclear translocation, we analyzed VSMCs by fluorescence microscopy and the depth of the magenta color illustrated the degree of p65 translocation. Our results revealed that LPS significantly increased nuclear translocation of NF-κB p65 which could be prevented by LEE treatment (Fig. 6).

Figure 6: Effect of LEE on nuclear accumulation of p65 induced by LPS in VSMCsConfluent VSMCs monolayers were grown on coverslips followed by being incubated with different conditions in the 24-hole plates for 48 h. Then cells were fixed, permeabilized, and stained with anti-p65 antibody and a secondary antibody conjugated to fluorescent red. The DNA was stained using DAPI. The negative control (p65 antibody not added) and a blank control are both included.

Mentions:
The activation of NF-κB contributes to inflammation status by promoting the expression of inflammation genes. To verify NF-κB p65 nuclear translocation, we analyzed VSMCs by fluorescence microscopy and the depth of the magenta color illustrated the degree of p65 translocation. Our results revealed that LPS significantly increased nuclear translocation of NF-κB p65 which could be prevented by LEE treatment (Fig. 6).

Aim:: Atherosclerosis is a kind of chronic inflammatory disease. A crucial pathology change of atherosclerosis is the migration of activated VSMCs to the intima where they interact with leukocytes by expressing adhesion molecules, including intercellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). Moreover, monocyte chemoattractant protein-1 (MCP-1) expressed by VSMCs plays an important role in recruiting monocytes and macrophages. Leech (Whitmania pigra Whitman) is a traditional Chinese medicine to treat cardiovascular diseases including atherosclerosis, however previous research has rarely reported the molecular mechanism for its curative effect. Thus, our study focuses on the effects of leech extracts on the expression of inflammatory factors, adhesion molecules and MCP-1 in rat VSMCs.

Results:: LEE suppressed LPS-induced VSMCs migration and decreased the chemotaxis and adhesive capacity of THP-1 and RAW264.7 to LPS-stimulated VSMCs. LEE also attenuated the upregulation of a variety of pro-atherosclerotic factors by inhibiting the phosphorylation of p38 MAPK. LEE was also observed to prevent NF-&kappa;B p65 nuclear localization using immune-fluorescent staining.